Uses of Growth Hormone Fragments
A growth hormone, human growth hormone technology, applied in the field of growth hormone peptide fragments, can solve problems such as limited clinical application
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Embodiment 1
[0169] Example 1: Determination of the effect of AOD on cartilage repair by assessing tissue formation:
[0170] method
[0171] Cell Culture and Therapy
[0172] Chondrocytes were aseptically isolated from articular cartilage obtained from the calf metacarpal-carpal joint and seeded on type II collagen-coated membrane inserts (Millipore TM )(1.5×10 6 cells / 12mm diameter membrane). Chondrocytes were grown in Ham's F-12 supplemented with 5% fetal bovine serum (FBS) under standard culture conditions. After 5 days, serum increased to 20%. Chondrocytes were then grown for 2 weeks in the presence and absence of AOD9604 (SEQ ID NO: 1 ) (10 or 100 μg / ml) (total culture time 3 weeks). Add fresh AOD each time the medium is changed.
[0173] organizational assessment
[0174] The tissue was harvested and digested with papain (Sigma, 40 g / ml) for 48 h at 65°C. DNA content was determined using Hoechst 33258 dye binding assay (Polysciences) and fluorometry (excitation 365 nm, emiss...
Embodiment 2
[0181] Example 2: Effect of AOD on natural cartilage
[0182] method
[0183] Cartilage explants were removed from the calf metacarpal-carpal joint and cultured in Ham's F-12 supplemented with 20% fetal bovine serum (FBS) in the presence or absence of AOD9604 (SEQ ID NO: 1) (AOD9604 was 10 or 100g / ml) for 1 week or (AOD9604 is 100 or 500g / ml) for 2 weeks.
[0184] organizational assessment
[0185] The tissue was harvested, and the water content and dry weight were measured. Tissues were then digested with papain (Sigma, 40 μg / ml) at 65°C for 48h. DNA content was determined using Hoechst 33258 dye binding assay (Polysciences) and fluorometry (excitation 365 nm, emission 458 nm). Proteoglycan content was determined by measuring the amount of sulfated glycosaminoglycans using a dimethylmethylene blue dye binding assay and spectrophotometry (525 nm). Collagen content was determined by measuring the hydroxyproline content of an aliquot of the hydrolyzed papain digest in 6NHCl a...
Embodiment 3
[0195] Example 3: Effect of peptide AOD9604 on the differentiation of myoblasts into myocytes in vitro
[0196] method
[0197] Cell Culture and Therapy
[0198] Under standard cell culture conditions, C2C12 was grown in monolayer culture in DMEM (high glucose) supplemented with 10% fetal bovine serum. Cells were passaged at 60-70% confluence. By making cells (3000 cells / cm 2 ) were grown for 3 days in the absence or presence of AOD9604 (10 and 100 μg / ml) to determine the effect of AOD9604 (SEQ ID NO: 1 ) on cell proliferation. DNA content was then quantified. Cells were digested by papain and DNA content was determined using Hoechst 33258 dye binding assay (Polysciences) and fluorometry (excitation 365 nm, emission 458 nm).
[0199] Cellular Differentiation Assessment
[0200] C2C12 (2×10 3 cells / cm 2 ) were grown in DMEM and 10% FBS. Differentiation was induced by replacing the growth medium with a differentiation medium consisting of DMEM supplemented with 2% hor...
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